Interleukin 21 (IL-21) medicine for cancer therapy
An interleukin and tumor technology, applied in anti-tumor drugs, drug combinations, pharmaceutical formulations, etc., can solve problems such as unsatisfactory tumor treatment effects, and achieve the effects of preventing tumor recurrence, improving microenvironment, and enhancing curative effects.
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Embodiment 1
[0057] Example 1. Intratumoral injection of IL-21 significantly enhanced the tumor therapeutic effect of anti-HER2 / neu antibody:
[0058] to normal wild-type Balb / c mice ( figure 1 A) or HER2 / neu antigen immunotolerant (HER2 / neu x Balb / c) F1 mice ( figure 1 B) Inoculate the tumor, and give anti-HER2 / neu antibody treatment when the tumor grows to a certain size, and at the same time inject recombinant IL-21 protein (Pepro Tech) into the tumor for combined treatment, which can significantly enhance the tumor therapeutic effect of anti-HER2 / neu antibody.
[0059] Specific experimental methods:
[0060] (1) Inoculate normal Balb / c mice subcutaneously with 8×10 5 Tubo tumor cells were divided into 4 groups, 5 mice in each group, anti-HER2 / neu antibody+IL-21 group: 100 μg anti-HER2 / neu antibody was intraperitoneally injected on day 16, and tumor cells were injected on day 16 and day 19. Intratumoral injection of 5 μg IL-21 protein (Pepro Tech); anti-HER2 / neu antibody group: intra...
Embodiment 2
[0062] Example 2. Intratumoral injection of IL-21 induces the polarization of tumor-associated macrophages to the M1 type, significantly improving the tumor immunosuppressive microenvironment:
[0063] Intratumoral injection of IL-21 into tumor-bearing mice significantly reduced the proportion of macrophages in tumor tissue ( figure 2 A), increasing anti-tumor effector cells (NK, CD8 + The proportion of T cells) ( figure 2 B) Induced tumor infiltrating macrophages with low expression of M2 macrophage factors CCL17, IL-10, TGF-β, VEGF, etc., and high expression of M1 type macrophage factors CXCL9, Nos2, etc. ( figure 2 C); Eliminate the inhibitory effect of TAM on T cell proliferation ( figure 2 D).
[0064] Specific experimental methods:
[0065] (1) Analysis of cell subsets in tumor tissue: Balb / c mice were subcutaneously inoculated with 8×10 5 Tubo tumor cells were injected with 10 μg of IL-21 protein (Pepro Tech) into the tumor on the 15th day after inoculation. Af...
Embodiment 3
[0074] Example 3. IL-21 directly induces macrophages to differentiate into M1 type:
[0075] In order to prove that the intratumoral injection of IL-21 induces the polarization of tumor-associated macrophages to the M1 type is the result of the direct effect of IL-21 on macrophages, we first demonstrated that intraperitoneal injection of IL-21 in mice can also significantly inhibit peritoneal macrophages. The cells expressed M2 macrophage factors CCL17, IL-10, TGF-β, Mrc-1 ( image 3 A); On this basis, I isolated mouse peritoneal macrophages and co-cultured with IL-21 in vitro, and cultured overnight can significantly inhibit the expression of peritoneal macrophages M2 macrophage factors VEGF and TGF-β ( image 3 B); In addition, IL-21 can also significantly increase the secretion of inflammatory cytokines TNF-β and IL-6 secreted by LPS-stimulated mouse peritoneal macrophages in vitro ( image 3 C); IL-21 directly acts on M2 tumor-infiltrating macrophages and can rapidly indu...
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